Sulbactam is a plausible option for treating Acinetobacter infection because of its intrinsic antibacterial activity against the Acinetobacter genus, but the mechanisms of sulbactam resistance have not been studied fully in Acinetobacter baumannii In this study, a total of 2197 clinical A. baumannii isolates were collected from 27 provinces in China. Eighty-eight isolates with various minimal inhibitory concentrations (MICs) for sulbactam were selected based on their diverse clonality and underwent multilocus sequence typing, antimicrobial-susceptibility testing and resistance gene screening. The copy number and relative expression of blaTEM-1D and ampC were measured via qPCR and qRT-PCR, respectively. The genetic structure of multicopy blaTEM-1D was determined using whole-genome sequencing technology. The cefoperazone/sulbactam resistance rate of the 2197 isolates was 39.7%. The positive rate of blaTEM-1D or ISAba1-ampC in the sulbactam-nonsusceptible group was significantly higher than that in the sulbactam-susceptible group (64.91% vs. 0%, P<0.001; 78.95% vs. 0%, P<0.001, respectively). The MIC of sulbactam (P<0.001) varied considerably between the ampC groups with or without upstream ISAba1 Notably, the genetic structure of the multicopy blaTEM-1D gene in strain ZS3 revealed that blaTEM-1D was embedded within four tandem copies of the cassette (IS26-blaTEM-1D-Tn3-IS26). Therefore, blaTEM-1D and ISAba1-ampC represent the prevalent mechanism underlying sulbactam resistance in clinical A. baumannii isolates in China. The structure of four tandem copies of blaTEM-1D firstly identified may increase sulbactam resistance.